Infertility is defined by most authorities as the inability to achieve a pregnancy after one year of unprotected intercourse. Conception is normally achieved within 12 months in 80-85 percent of couples using no contraceptive measures; thus an estimated 15 percent of couples attempting their first pregnancy will have difficulty conceiving.
Grape Seed and Endometriosis
Resveratrol, a potent isoflavone found in grape seed, is best known for its potential life extending properties as shown by studies in Drosophila Megaloblaster (fruit fly) and budding yeast, Saccharomyces cerevisiae. Canton-S flies cultured on food containing 200 M resveratrol showed an increase in average lifespan of 17% for females and 10% for males and an increase in maximum lifespan of 22% and 20% for females and males, respectively.
Although it is known to exert pleiotropic effects the lifespan extending effect of resveratrol is thought to be mediated by means of the activation of the histone deacetylase Sir2. Sir2 has been implicated in lifespan extension of yeast, nematodes, and fruit flies, and it has been proposed to mediate some of the effects of calorie restriction. Consistent with the importance of Sir2 activity in lifespan determination, scientific data showed that treatment of flies with the Sir2 inhibitor sirtinol leads to shortening of lifespan Bauer, et a., 101(35) Proc Natl Acad Sci USA 12980-12985 (2004).
Resveratrol has also been shown to protect ovarian follicles be antagonizing the aryl hydrocarbon receptor gene that sensitizes them to toxic polycyclic aromatic hydrocarbons (PAHs). Researchers provided molecular evidence for a pathway involved in the elimination of the female germline due to prepregnancy and/or lactational exposure to (PAHs), environmental toxicants found in cigarette smoke. Ovaries of offspring born to mice exposed to PAHs contained only a third of the ovarian follicle pool compared with offspring of unexposed female mice. Activation of the cell death pathway in immature follicles of exposed females was mediated by the aryl hydrocarbon receptor (Ahr), as ovarian reserve was fully rescued by maternal cotreatment with the Ahr antagonist, resveratrol, or by inactivation of the Ahr gene. Furthermore, in response to PAHs, Ahr-mediated activation of the harakiri, BCL2 interacting protein (contains only BH3 domain), was necessary for execution of cell death. Jurisicova, et al 117(12) J Clin Invest 117(12):3971-8 (2007).
One study found, however, that grape seed extract increased IL-6 levels in cultured neurons. Grape polyphenols are known to protect neurons against oxidative stress. Grape seed extract (GSE) from “Koshu” grapes (Vitis vinifera) containing a variety of polyphenols, was subjected transcriptome analysis to determine the effects of GSE on primary cultures of astrocytes in the hippocampus. GSE upregulated various mRNAs for cytokines, among which interleukin-6 (IL-6) showed the biggest increase after treatment with GSE. The GSE-evoked increase in IL-6 mRNAs was confirmed by quantitative RT-PCR. IL-6 proteins were detected by ELISA in the supernatant of GSE-treated astrocytes and an oxidative stress-induced neuronal cell death model in vitro using a neuron rich culture of the hippocampus was developed. Treatment of the neurons with H(2)O(2) caused neuronal cell death in a time- and concentration-dependent manner. Exogenously applied IL-6 protected against the H(2)O(2)-induced neuronal cell death, which was mimicked by endogenous IL-6 produced by GSE-treated astrocytes. Taken together, GSE acting on astrocytes increased IL-6 production, which functions as a neuroprotective paracrine, could protect neuronal cells from death by oxidative stress. Fujishita, K, et al, Cell Mol. Neurobiol. 2009 Apr. 21. [Epub ahead of print].
The majority of research shows the contrary. Over recent years, resveratrol, found in grape seed, has been shown to inhibit levels of Interleukin-6 (IL-6) associated with infertility particularly that resulting from endometriosis. Endometriosis is a benign estrogen-dependent gynecological disease, which affects approximately 10% of women of reproductive age, is characterized by the presence of endometrial tissue outside the uterine cavity, and is associated with pelvic pain, dysmenorrea and infertility. Despite all the research accumulated over the years, the etiology and the pathogenesis of endometriosis are still unclear. Minici, et al, 23(3) Human Reproduction 23(3):530-537 (2008). It has been found in the majority of women with a regular menstrual cycle and infertility. Kunz, G, et al, 20(8) Hum Reprod: 2309-16 (2005).
Kunz, et al, Id., studied 227 women with regular menstrual cycles (mean 29 days, range 21-28) aged 17-46 years (mean 32.5) each after giving informed consent. Together with their male partners, they had a history of infertility prompting them to have a sterility work-up and subsequent treatment in Dr. Kunz' infertility clinic. Of these patients, 160 women (aged 17-46 years; mean 32.3) with a history of infertility of 1-13 years (mean 3.6) were suffering from endometriosis as demonstrated by laparoscopy. Almost one half of these patients presented with minimal or mild endometriosis (n=81) and the rest with moderate or severe endometriosis (n=79), according to the revised classification of the American Society of Reproductive Medicine (ASRM). No additional factors responsible for their female infertility could be identified.
Endometriosis is so highly associated with elevated levels of the inflammatory cyokine that elevated levels of IL-6 in women with endometriosis is used as reliable non-invasive marker for the disease. A serum IL-6 threshold of 25.75 pg/ml afforded a sensitivity of 75% and specificity of 83% in the diagnosis of MM endometriosis. Sensitivity and specificity for CA-125 in the diagnosis of MS endometriosis, using 35 IU/ml as the cut-off value, were 47% and 97%, respectively. Martinez, S et al, 22(3) Hum Reprod 836-42 (2007).
The prevalence of adenomyotic lesions in all 160 women with endometriosis was 79%. In women with endometriosis below an age of 36 years and fertile partners, the prevalence of adenomyosis was 90% (P<0.01) With a prevalence of up to 90%, uterine adenomyosis is significantly associated with pelvic endometriosis and constitutes an important factor of sterility in endometriosis presumably by impairing uterine sperm transport. Kunz, G, et al, 20(8) Human Reproduction 2309-2316 (2005)
Even in the absence on adenomyotic lesions, Interleukin-6 (IL-6) that has a negative impact on sperm motility. Even in the absence on adenomyotic lesions, Interleukin-6 (IL-6) that has a negative impact on sperm motility. The expression of IL-6 and its receptor was detected in human granulosa cancer cells. Estradiol secretion was significantly inhibited by adding IL-6, which also suppressed aromatase activity to 50% of the control. The findings demonstrated that IL-6 may reduce estrogen production in human granulosa cells and may support the notion that IL-6 is related to impaired estrogen biosynthesis in patients with endometriosis. Deura, et al, 83 Suppl 1 Fertil Steril 1086-92 (2005).
Other grape polymers have found to have similar activities in modulating IL-6. Researchers examined the potential antioxidant activity and the immunopharmacological activity of new epicatechin conjugates obtained by depolymerization of grape polymeric flavanols in the presence of cysteamine or cysteine and with or without gallate. The compounds studied were (−)-epicatechin (1), cysteinyl-epicatechin (2), cysteamine-epicatechin (3), (−)-epicatechin gallate (4), cysteinyl-epicatechin gallate (5), and cysteamine-epicatechin gallate (6) When incubated with an erythrocyte suspension, flavanols protected the erythrocyte membrane from hemolysis induced by 2,2′-azobis(2-amidinopropane)dihydrochloride, an azo free-radical initiator. All the epicatechin derivatives tested were more efficient as antioxidant than epicatechin. The compounds were tested for their capacity to modulate IL-1beta and IL-6, which are the main cytokine factors influencing the acute phase of the inflammatory response. (−)-Epicatechin and its related compounds inhibited the production of IL-1beta and IL-6 in whole blood incubated in the presence of Escherichia coli lipopolysaccharide. Mitjans, M, et al, 52(24) J Agric Food Chem 7297-9 (2004).
Other researchers using transvaginal ultrasound examinations measured serum IL-6 concentrations in 14 patients with ovarian endometrioma and 4 patients with benign gynecologic disease without endometriosis. IL-6 was significantly higher in patients with endometrioma than in those without endometriosis at the time of diagnosis. The mean serum IL-6 concentration significantly decreased after the operation for the disease. Yoshida, S, et al, 54 Suppl 1 Gynecol Obstet Invest 24-7; discussion 27-9 (2002).
Detailed studies have demonstrated the effects of various other types of grapes on IL-6 in the context of the mast cell-mediated allergy model in vitro. Kim, S, Kwoh, T, and Shin, T, 233 Experimental Biology and Medicine 192-199 (2008). In the Kim study, a preparation of Methanol Extract of V. amurensis (MEVA) was prepared and assays performed for its effect on TNF-{alpha}, IL-6, and IL-8 Secretion. TNF-{alpha}, IL-6, and IL-8 secretion was measured by modification of an enzyme-linked immunoabsorbent assay (ELISA). Cultured cells treated with the extract showed dose-dependently inhibited the secretion of TNF-{alpha}, IL-6, and IL-8 in PMACI-stimulated HMC-1 cells.
Similar findings were observed in animals. The inhibition pattern on the release of IL-6 was similar to that seen with TNF-alpha. In a related in vivo study, rats were fed a diet containing 5% (wt/wt) dried muscadine grape skins (Vitis rotundifolia) for 14 days and then were injected with carrageenan in the foot pad. After 3 h, paw edema was measured and the rats on the grape skin diet had approximately 50% less paw edema than controls (P<0.05). These results demonstrate that the muscadine grape skin powder possesses significant in vitro and in vivo antiinflammatory properties. Greenspan, et al 53(22) J Agric Food Chem 8481-4 (2005).
Para-aminobenzoic Acid (PABA) and Folate Levels
In addition to the well-established association between folic acid deficiency during pregnancy and neural tube and other birth defects, Wolff, T, et al, 150(9) Ann Intern Med 632-9 (2009), the intake of folic acid supplements, with or without additional multivitamins, is associated with a reduction in infertility disorder due to ovulatory disorder. Chavarro J E, et al, 89(3) Fertil Steril 89(3):668-76 (2008). Chavarro, et al, studied 438 women with reported infertility due to ovulatory disorder for 8 years. There was an inverse association between frequency of multivitamin use and ovulatory infertility. The multivariate-adjusted relative risk (95% confidence interval) of ovulatory infertility was 0.88 (0.60, 1.28) for women consuming 2 tablets/week or less, 0.69 (0.51, 0.95) for women consuming 3 to 5 tablets/week and 0.59 (0.46, 0.75) for women consuming 6 or more tablets/week, when compared to women who did not use these supplements (P, trend<0.001). Folic acid appeared to explain part of the association between multivitamin supplement use and risk of ovulatory infertility. Adjusting for multi-vitamin use attenuated, but did not vitiate, the association with folic acid intake alone. The researchers noted that supplementation with folic acid by itself or as part of a multivitamin has been shown to reduce the risk of neural tube defects and may prevent other congenital malformations, and suggested that women planning to become pregnant should consider taking a multivitamin as this may also help them to become a pregnant.
Reduced levels of folic acid or folate and increased homocysteine are associated with infertility. Possible mechanisms of the deleterious effects of folate deficiency and homocysteine accumulation on female fertility include, as in the male, reduced cell division (e.g. of oogonia during oogenesis or of granulosa cells during folliculogenesis), inflammatory cytokine production, altered NO metabolism, oxidative stress, apoptosis and defective methylation reactions. Forges, T, et al, 13(3) Hum Reprod Update 225-38 (2007).
While opposite results have been observed by researchers, Avivia and Boxtel, 11(6) Trop Med Int Health 11(6):804-8 (2006), there are several reports of increased folic acid turnover during pregnancy and a resulting deficit. Gregory, J F, et al, J Nutr 131(7):1928-37 (2001); Higgins, J R, et al, 107(9) BJOG 107(9):1149-54 (2000). Para-amino benzoic acid (PABA) is metabolized into folic acid both by polymorphonuclear neutrophils, Sagone A L Jr, Husney R M & Davis W B 14(1) Free Radic Biol Med 27-35 (1993) and gut bacteria, Camilo E, et al, 110(4) Gastroenterology 991-8 (1996). Hence, the supplementation of PABA in addition to the standard folic acid supplementation provides a pool of folic acid precursor that may be utilized to replenish catabolized folic acid.
Red Clover and Phytoestrogens
The need for adequate levels of estradiol to conceive and maintain pregnancy is well-established, and serves as the basis for several of the pharmacological treatments for infertility. Ming-fang Miao and He-feng Huan, 10(1) J Zhejiang Univ Sci B 10(1): 35-45 (2009). The isoflavone genistein GEN is a saponin found in soy (Glycine max) and red clover (Trifolium pratense). The estrogenic activity of GEN is known, and it is widely advertised as a phytoestrogen useful in alleviating climacteric complaints and other postmenopausal disorders. Knowledge of effects of long-term administration of GEN in laboratory animals is scarce, and effects in the uterus and mammary gland after long-term administration have not been studied. The uterus and mammary gland are known to be negatively influenced by estrogens used in pharmaceutical hormone therapy. In rats treated with either of estrogen doses or the high GEN doses, researchers found increased uterine weight, and histologic analysis showed estrogen-induced features in the uteri. In vaginae, these same treatments induced hyperplastic epithelium compared with the atrophic controls. The endometrium was composed of cuboidal inactive cells, and the connective tissue was an unorganized lax syncytium of round nuclei. No mitotic activity was detected in epithelial cells. Serum levels of luteinizing hormone were decreased by estrogen but not by GEN. Rimoldi, et al, 115 (S-1) Environ Health Perspect 62-68 (2007).
The relative oestrogenic activities of 0.1 g each of extracts of phytosterol, soy isoflavone, red clover, kudzu and soybean extracts were determined using this assay and found to be equivalent to 212, 1, 3.2, 132 and 1025 nM of 17 beta-estradiol, respectively. Bennetau-Pelissero, et al C 15(1) Phytochem Analysis 40-5 (2004).
Vitex agnus-castus and Luteal Phase Progesterone
Low levels of progesterone can result in infertility along with short menstrual cycles as the progesterone is required to maintain the corpus luteum prior to fertilization. Chimpanzees in Gombe National Park consume fruits of Vitex fischeri during a short annual fruiting season. This fruit species is a member of a genus widely studied for phytoestrogen composition and varied physiological effects. One particularly well-studied species, V. agnus-castus, also known as chaste-berry, is noted for its documented effects on female reproductive function evidenced in increased progesterone levels and consequent regulation of luteal function. Thompson, E M, 70(11) Am J. Primatol. 2008 November; 70(11):1064-71. (2008)
Results of a small clinical trial involving 96 women with fertility disorders (38 women with secondary amenorrhea, 31 with luteal insufficiency, and 27 with idiopathic infertility) suggested that patients receiving chasteberry achieved pregnancy more readily than did women in the placebo group. The subjects received chasteberry or placebo twice daily for three months. Hormone levels did not differ, but in women with amenorrhea or luteal insufficiency, pregnancy occurred in the active treatment group more than twice as often as in the group receiving placebo. However, the total number of patients conceiving was small (15 women), the treatment was only administered for three months, and the product used contained five additional herbs. Roemheld-Hamm, B, 72 Am Fam Physician 821-4(2005).
In another small study involving 52 patients with luteal phase defects, women in the active treatment group were found to have reduced prolactin release, normalized luteal phases, improved luteal phase progesterone synthesis, and increased luteal phase estradiol. This study appeared to support the use of chasteberry for luteal phase disorders, but the actual effect on fertility was not mentioned. A recent double-blind placebo-controlled pilot study 27 of 30 women showed an increasing trend in midluteal phase progesterone level and an increased number of pregnancies in the 15 women who took a nutritional supplement containing chasteberry for five months. Roemheld-Hamm, B, 72 Am Fam Physician 821-4 (2005).
Even in wild chimpanzees, hyperprogesteronemia has been associated with Vitex fischeri consumption. Chimpanzees in Gombe National Park consume fruits of Vitex fischeri during a short annual fruiting season. This fruit species is a member of a genus widely studied for phytoestrogen composition and varied physiological effects. One particularly well-studied species, V. agnus-castus, is noted for its documented effects on female reproductive function, evidenced in increased progesterone levels and consequent regulation of luteal function. Researchers examined reproductive hormone levels in both male and female chimpanzees during a 6-week period of intense V. fischeri consumption. V. fischeri consumption was associated with an abrupt and dramatic increase in urinary progesterone levels of female chimpanzees to levels far exceeding the normal range of variation. Female estrogen levels were not significantly impacted, nor were male testosterone levels. Thompson, E M, 33(4) Clin Exp Obstet Gynecol 205-8 (2006).
An Agnus castus-containing homeopathic preparation was investigated in Germany in 67 women with fertility disorders who received 50 drops of Phyto Hypophyson L or placebo 3 times a day over 3 months or 3 cycles. The outcome measure were spontaneous menstruation, improved concentration of progesterone in the luteal phase, shortening of the cycle, and earlier ovulation. Pregnancy was achieved in 38 out of 67 women, more often from women with oligomenorrhea in the Phyto Hypophyson L group compared to the placebo group (82 versus 45%, p=0.021). However, there was no significant effect when viewing the whole group. The baby take-home rate during therapy and 6 months after the end of therapy showed a ratio of 6:2 (18.7:6.4%), which was not significant. Furthermore, in the oligomenorrhea verum group we observed a significant increase of progesterone during the luteal phase compared to the oligomenorrhea placebo group. Bergmann, 7(4) Forsch Komplementarmed Klass Naturheilkd. 190-9 (2000).
In another German study of another Vitex agnus castus preparation and placebo, subjects were given 20 mg of either the Vitex agnus-castus or placebo. Aim of the study was to prove whether the elevated pituitary prolactin reserve can be reduced and deficits in luteal phase length and luteal phase progesterone synthesis be normalized. Blood for hormonal analysis was taken at days 5-8 and day 20 of the menstrual cycle before and after three month of therapy. Latent hyperprolactinaemia was analysed by monitoring the prolactin release 15 and 30 min after i.v. injection of 200 micrograms TRH. 37 complete case reports (placebo: n=20, Vitex n=17) after 3 month of therapy were statistically evaluated. The prolactin release was reduced after 3 months, shortened luteal phases were normalised and deficits in the luteal progesterone synthesis were eliminated. These changes were significant and occurred only in the group treated with Vitex agnus-castus. Milewicz, A, 43(7) Arzneimittelforschung 752-6 (1993).
Eleuthero senticosus and phytoestrogens
There are a group of so-called tonic remedies in Far Eastern medicine which are traditionally viewed as harmonizing or adjustive. Ginseng and eleutherococcus are the best known, and there is evidence that they increase arousal, stamina and stress resistance. We have attempted to explore the relationship between the behavioral and the stress effects, and to relate this to traditional concepts. In one series of experiments mice were given ginseng throughout their lifespan. At intervals their behavior response to mild stress was examined and found to be exaggerated compared to controls without ginseng. However, normal ambulatory behavior in the absence of stress was unaffected. A second series of experiments indicated that the binding of corticosteroid to certain brain regions was increased in adrenalectomized rats given ginseng saponin, compared to saline treated controls. This can be interpreted as a result of an increase in hypothalamic-pituitary-adrenal sensitivity caused by ginseng saponin. This is in accord with traditional concepts of the use of these remedies. Fulder S J, 9(2) Am J Clin Med 112-8 (1981).
Eleuthero senticosis, also known as ginseng, has been shown to attenuate nerve grown factor, associated with polycystic ovaries, in female rats. Polycystic ovary was fully developed in rats with a single intramuscular injection of estradiol valerate. Increased expression of nerve growth factor (NGF) was noted in the ovaries and the brain of rats with polycystic ovaries. Korean red ginseng total saponins administration attenuated NGF expression in the ovaries but not in the brain. Pak, et al 84 Supp 2 Fertil Steril 1139-43 (2005). Ginseng has also been shown to produce increased embryonic growth at 50 micrograms/ml and hindbrain, midbrain and caudal neural tube at 100 micrograms/ml with Ginsenoside Rc and the pGPx mRNA level increased at all doses. Lee, et al, 54(3) J Reprod Devel 164-70 (2008).
Eleuthrococcus has also been shown to protect the developing embryo. Preventive administration of Eleutherococcus extract during prenatal and pre-embryonic periods of development prevents embryotoxic effect of subsequent treatment of pregnant rats with ethanol and sodium salicylate. Eleutherococcus abolishes embryotoxic and teratogenic effects of ethanol manifested against the background of experimental syndrome of iron deficit in pregnant females. Mechanism of its antiteratogenic action is probably based on stimulation of cell detoxification mechanisms, increase in energy potential of cells, as well as on stabilization of structural and functional state of cell membranes. Gordeichuk T, et al, 24(1) Ontogenez. 1993 48-55 (1993)
Panax ginseng and Eleutherococcus senticosus may exaggerate an already existing biphasic response to stress via inhibition of enzymes which limit the binding of stress hormones to their receptors. Specifically, it is suggested that PG inhibits 11-beta hydroxysteroid dehydrogenase one and ES inhibits catechol-O-methyl transferase, both of which reside in close proximity to stress hormone receptors and catalyse the degradation of stress hormones into inactive compounds. In addition, it is suggested that the increased energy said to result from PG and ES may be a consequence of their occupancy of stress hormone receptors. Gaffney B T, Hugel, H M, and Rich, P A, 56(5) Med Hypothesis 567-72 (2001)
Nutritional therapies have included Vitex agnus-castus, alpha lipoic acid, glucosamine, folic acid, and standard multiple vitamins.
Only a few published teachings claim to affect this serious and prevalent problem in the population through intervention in the nutritional status of patients. U.S. Patent Application No., 20070104801, to Cecchi, teaches an undescribed fertility enhancing and anti-aging composition that may contain any of alpha-lipoic acid, glucosamine sulfate or Peruvian Maca with or without horny goat weed.
U.S. Pat. No. 7,115,650, to Broqua, teaches a method of improving fertility by the administration of an inhibitor of dipeptidyl peptidase IV, wherein said inhibitor is an amino-acyl pyrrolidine or an amino-acyl thiazolidine.
U.S. Pat. No. 6,569,857, to Hermelin, teaches a method for increasing the possibility of conception while enhancing nutritional stores for a developing embryo or fetus prior to and during pregnancy by administering to an animal during a period commencing prior to at least two weeks before conception a specific dose of vitamin B6 and folic acid.
U.S. Pat. No. 7,410,657, to Lee, teaches several compositions for enhancing fertility that require a large number of herbs used in Chinese medicine, including Radix Polygalae Tenuifoliae, Semen Ziziphi Spinosae, Radix Ginseng, Rhizoma Atractylodis Macrocephalae, Rhizoma Zingiberis Officinalis Recens, Sclerotium Poriae Cocos, Radix Astragali, Radix Ligustici Wallichii, Radix Angelicae Sinensis, and the like.
U.S. Pat. No. 6,989,164, to Trant, teaches a pharmaceutical composition for oral ingestion having fertility promoting activity in males comprising components present in the proportion in parts by weight of: about 5 to 50% green tea, vitamin C, vitamin E, and selenium, about 10 to 80% L-carnitine, up to about 1% ferulic acid in Dong quai, up to about 1% vitamins B6, B12, and folate, up to about 10% zinc. The combination of L-carnitine and Coenzyme Q10, but not Lepidium Meyenii, is taught by another patent to Trant, U.S. Pat. No. 7,045,151 for a pharmaceutical composition having fertility promoting activity in males comprising components present in the proportion in parts by weight of about 20 to 80% L-carnitine, about 5 to 30% vitamins C and E, about 0.1 to 10% coenzyme Q10, about 0.001 to 1% selenium, about 0.2 to 20% ferulic acid, about 0.1 to 2% zinc, and about 0.001 to 1% B vitamins.
U.S. Pat. No. 6,861,079, to Sweazy, teaches a fertility kit to enhance natural fertility and detect female ovulation and male fertility potential comprising specific dose ranges of L-Arginine, L-Cysteine, Selenium, Vitamin C, Vitamin E, Zinc, Astragalus, Pycnogenol, Vitamin B-6, Para-aminobenzoic acid (PABA), Vitamin A, Folic Acid, at least one phytoestrogen, along with several devices used in the promotion of ovulation timing and intercourse.
U.S. Pat. No. 6,372,266, to Sweezey, teaches the use of resveratrol in infertility. U.S. Patent Application No. 20070099826, to Wong teaches a compound capable of modulating transcription arising from an egr-1 response element consensus sequence and expression state of a gene in manufacture of a medicament for the treatment of a disease or health condition associated with an expression state of a gene associated with an egr-1 response element consensus sequence that may be resveratrol and may be used to treat infertility. Neither of these patents teach the combination of resveratrol with para amino-benzoic acid and red clover extract.
This inventor has discovered the elegant combination of these three nutrients, each with a primary effect on a different aspect of process of conception and pregnancy